title: Prostaglandin F2 alpha receptors in the human trabecular meshwork.
abstract: OBJECTIVE
  Prostaglandin F2 alpha (PGF2 alpha) and analogs, such as latanoprost, are thought to lower intraocular pressure (IOP), primarily by increasing uveoscleral outflow. However, outflow through the trabecular meshwork may be increased as well. The authors hypothesize that any effect on the trabecular meshwork is mediated by prostanoid FP receptors (receptors for prostaglandin F2 alpha) in this tissue.
  
  
  METHODS
  To test this hypothesis, tissue sections of the human trabecular meshwork and cultures of human trabecular meshwork cells were examined for the presence of FP receptors using immunofluorescence microscopy with affinity-purified antibodies raised against a glutathione-S-transferase (GST)-FPA receptor fusion protein. The presence of the receptor was confirmed by using reverse transcription-polymerase chain reaction (RT-PCR), functional assays of PGF2 alpha-stimulated inositol phosphate hydrolysis, and intracellular calcium measurements.
  
  
  RESULTS
  Positive FPA receptor immunolabeling was observed in sections of the human trabecular meshwork and in cultured human trabecular meshwork cells. In both cases, specific labeling could be blocked by preincubation with a GST-FPA receptor fusion protein. Cross-blocking experiments with other receptor fusion proteins did not block specific labeling in cultured trabecular meshwork cells. PGF2 alpha caused a dose-dependent increase in total inositol phosphate accumulation and intracellular calcium release in human trabecular meshwork cells that was consistent with the presence of FP receptors. Using RT-PCR, message-encoding prostanoid FPA receptors were found in total RNA isolated from human trabecular meshwork cells.
  
  
  CONCLUSIONS
  Prostanoid FPA receptors exist in human trabecular meshwork cells, as shown by the presence of mRNA, protein, and functional response to PGF2 alpha. This study indicates that functional FP receptors are present in the human trabecular meshwork and that they may be involved in mediating some of the IOP-lowering effects of PGF2 alpha in the eye.
authors.affiliation: Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona, Tucson 85721, USA.
authors.forename: T L
authors.initials: TL
authors.lastname: Anthony
authors.forename: K L
authors.initials: KL
authors.lastname: Pierce
authors.forename: W D
authors.initials: WD
authors.lastname: Stamer
authors.forename: J W
authors.initials: JW
authors.lastname: Regan
mesh_terms: D000818:Animals; D019556:COS Cells; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D002645:Chickens; D017931:DNA Primers / Q000737:chemistry; D015237:Dinoprost / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D006868:Hydrolysis; D007295:Inositol Phosphates / Q000378:metabolism; D018613:Microscopy, Confocal; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D011982:Receptors, Prostaglandin / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014129:Trabecular Meshwork / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection
chemical_list: D017931:DNA Primers; D007295:Inositol Phosphates; D012333:RNA, Messenger; D011982:Receptors, Prostaglandin; D011993:Recombinant Fusion Proteins; C034671:prostaglandin F2alpha receptor; D015237:Dinoprost; D002118:Calcium